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High-efficiency compound microbial inoculum for fermenting beds, and preparation method and application thereof

A compound bacterial agent and fermentation bed technology, applied in biochemical equipment and methods, methods based on microorganisms, bacteria, etc., can solve the problems of uneven technology, simple production process, and shortage, so as to improve feed utilization and economic benefits , the effect of increasing the stocking density

Active Publication Date: 2011-01-19
FEED RESEARCH INSTITUTE CHINESE ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] In recent years, with the rapid development of large-scale and intensive pig farming in China, the production efficiency has been greatly improved, but the traditional cement floor feeding mode has also produced some disadvantages. One is the concentrated discharge of pig manure, which requires a large area of ​​manure Farm and flushing water, wasting resources and labor, and causing serious pollution to the breeding environment and the surrounding environment. The annual production of livestock and poultry manure in my country has reached 1.9 billion tons, more than twice the amount of industrial solid waste discharge
The second is the toxic and side effects caused by the abuse of antibiotics, drug residues and bacterial resistance, and the quality and safety of livestock products; therefore, pig production must seek a new pig breeding production method that can reduce environmental pollution and improve pork food safety.
[0005] At present, my country is in the rising stage of the promotion and application of fermentation beds. Compared with the large demand in the breeding market, professional and efficient fermentation bed products and technologies are very scarce. In order to pursue profits, many fermentation bed products and technologies on the market vary. Some manufacturers directly collect indigenous Bacteria, packaged and sold after simple treatment, product quality and effect cannot be guaranteed
Some units have introduced compound micro-ecological preparations for raising pigs in fermented beds, but the existing problems are: the production process is too simple, the product quality and effect are average, and the method of preparing fermented bedding materials still needs to be fermented in advance, and the thickness of the bedding materials must be The length is 80-100cm, the operation process is complicated and wastes manpower and material resources
These have all limited the extensive popularization and application of fermentation bed

Method used

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  • High-efficiency compound microbial inoculum for fermenting beds, and preparation method and application thereof
  • High-efficiency compound microbial inoculum for fermenting beds, and preparation method and application thereof
  • High-efficiency compound microbial inoculum for fermenting beds, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1: slant strain activation

[0044] (1) Activation of the slant strain of Lactobacillus plantarum: take the ampoule tube strain, inoculate it into the test tube slant medium by aseptic operation, and cultivate it at a constant temperature of 37°C for 72 hours to prepare the test tube slant strain, and store it in the refrigerator at 4°C for later use. Slant medium (g / L): MRS medium: protein age: 10.0g, beef powder: 5.0g, yeast powder: 4.0g, glucose: 20.0g, Tween 80: 1.0mL, dipotassium hydrogen phosphate: 2.0g , sodium acetate: 5.0g, triammonium citrate: 2.0g, magnesium sulfate: 0.2g, manganese sulfate: 0.05g, distilled water: 1000mL, agar 15g, sterilized at 121°C for 20min.

[0045] (2) Activation of Candida tropicalis and Trichoderma konningen slant strains: Take ampoule tube strains and inoculate them into test tube slant medium by aseptic operation respectively, and culture at 30°C for 48 hours to make test tube slant strains, 4°C Store in the refrigerato...

Embodiment 2

[0047] Embodiment 2: the preparation of seed solution

[0048] (1) Preparation of Lactobacillus plantarum seed liquid: take the slant bacteria stored in the refrigerator, inoculate aseptically in a 500ml Erlenmeyer flask containing 200ml of MRS medium, and culture at 37°C and 150rpm for 20h. Seed medium: no agar was added to the slant medium in Example 1 (1) of the present invention.

[0049] (2) Preparation of Candida tropicalis and Trichoderma korningii seed liquid: take the slant strains stored in the refrigerator, inoculate them aseptically into 500ml triangular flasks with 100ml of wort medium respectively, and cultivate them at 30°C and 100rpm for 24h . Seed medium: no agar was added to the slant medium in Example 1 (2) of the present invention.

[0050] (3) Preparation of Bacillus subtilis seed solution: take the slant strains preserved in the refrigerator, inoculate them aseptically into 500 ml Erlenmeyer flasks containing 30 ml of LB medium, and culture at 35° C. an...

Embodiment 3

[0051] Embodiment 3: the preparation of high-efficiency composite bacterial agent

[0052] Inoculate Lactobacillus plantarum into a 50L fermenter with 30L fermentation medium at the same time according to the inoculation amount of 10%, and control the initial pH value to 6.5; the stirring speed is 100rpm, the ventilation rate is 0.15V / V min, and cultured at 30°C for 20h ; Simultaneously inoculate Candida tropicalis and Trichoderma korningii seed liquid according to 5% inoculation amount respectively again, stirring speed 100rpm, ventilation volume 0.3V / V min, 25 ℃ of cultivation 12h; Respectively simultaneously inoculate Bacillus subtilis again, The stirring speed is 200rpm, the ventilation rate is 0.8V / V min, and cultured at 37°C for 20h; the number of viable bacteria reaches 1×10 9 cfu / ml, 8×10 8 cfu / ml, 5×10 8 cfu / ml, 1×10 9 cfu / ml, 3×10 9 cfu / ml, stop the fermentation, sub-package and seal to obtain the compound microbial agent.

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Abstract

The invention provides a high-efficiency compound microbial inoculum for fermenting beds, particularly relates to a compound microbial inoculum for the fermenting beds, and a preparation method and application thereof. The compound microbial inoculum is composed of five beneficial bacterium strains (Bacillus subtilis CGMCC1.769 and CGMCC1.210, lactobacillus plantarum CICC6073, Candida tropicalis CGMCC2.587 and Trichoderma koningii CGMCC3.4290) and metabolites thereof (amylase, protease, lipase, cellulase, organic acid, bacteriophage and the like). The compound microbial inoculum is prepared by using a submerged liquid mixed fermentation technique. The method comprises the following steps: carrying out liquid submerged fermentation on the lactobacillus plantarum at 35 DEG C for 12 hours, respectively inoculating Candida tropicalis and Trichoderma koningii, carrying out mixed fermentation at 30 DEG C for 12 hours, inoculating Bacillus subtilis, carrying out liquid submerged mixed fermentation for 24 hours, respectively controlling the fermented viable count at 1-2*10<9> cfu / mL, 5-8*10<8> cfu / mL, 3-5*10<8> cfu / mL, 3-5*10<9> cfu / mL and 2-5*10<9> cfu / mL, finishing fermentation, packaging and sealing to obtain the high-efficiency compound microbial inoculum for the fermenting beds. The invention also provides application of the high-efficiency compound microbial inoculum-a preparation method of padding material. The compound microbial inoculum has the advantages of high viable count, high biological activity and low impurity bacterium pollution; the compound microbial inoculum can obviously reduce the odor of pigsties and reduce the living possibility of mosquitoes and flies, thereby achieving the ecological culture goals of no pollution and zero discharge; and the process for manufacturing the ferment bed is simple, thereby saving the padding material, human power and the like.

Description

technical field [0001] The invention relates to a high-efficiency composite bacterial agent for raising pigs in a fermentation bed, and also relates to a preparation method and application of the composite bacterial agent, belonging to the field of feed additives. Background technique [0002] In recent years, with the rapid development of large-scale and intensive pig farming in China, the production efficiency has been greatly improved, but the traditional cement floor feeding mode has also produced some disadvantages. One is the concentrated discharge of pig manure, which requires a large area of ​​manure Farm and flushing water, wasting resources and labor, and causing serious pollution to the breeding environment and the surrounding environment. The annual production of livestock and poultry manure in my country has reached 1.9 billion tons, more than twice the amount of industrial solid waste discharge. The second is the toxic and side effects caused by the abuse of ant...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/16C12N1/14A01K1/015C12R1/125C12R1/885C12R1/74C12R1/25
Inventor 王金全孙占敏张广民李红霞胡丽博陈鲜鑫王旭莉
Owner FEED RESEARCH INSTITUTE CHINESE ACADEMY OF AGRICULTURAL SCIENCES
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